For controls, the criteria and warnings provided previously for standard PCR for the detection of Nosema spp. are applicable (section For quantification, N. apis and N. ceranae recombinant amplicons should be used as external standards. Set up a standard curve using serial dilutions of recombinant target DNA fragments ranging from 10-2 to 10-8 and include them as quantification standards in every PCR run (see also the BEEBOOK paper on molecular methods (Evans et al., 2013)).